PB1337 - Native vascular extracellular matrix coating for in vitro cardiovascular models

Tuesday, June 25, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Co-Author(s)

TL

Titus P. Lemmens

PhD student
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands

Presenting Author(s)

Vanessa Bröker

PhD student
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands

Co-Author(s)

MR

Minke Rijpkema

PhD student
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands
SA

Salwan Al-Nasiry

Gynaecologist
2Department of Obstetrics and Gynaecology, GROW School for Oncology and Developmental Biology, Maastricht University
Maastricht, Limburg, Netherlands
CH

Christopher CW Hughes

Professor
3Department of Molecular Biology and Biochemistry, and Department of Biomedical Engineering, University of California, Irvine, USA
Irvine, California, United States
Rory R. Koenen

Professor
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands
Leon J. Schurgers, PhD

Professor
Department of BIochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands
JC

Judith MEM Cosemans

Associate Professor
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University
Maastricht, Limburg, Netherlands

Background: Extracellular matrices (ECM) are often neglected in developing models for studying cardiovascular diseases. The crosstalk between cells and matrix is crucial for vessel homeostasis and cellular phenotypes. Current in vitro models utilize singular matrix proteins or murine tumor-derived Matrigel® as culture coatings, which do not represent the in vivo situation.

Aims: Here, we aim to extract and characterize native human arterial matrix (HAM) from the umbilical cord to improve translatability of cardiovascular in vitro models.

Methods: HAM was produced from umbilical arteries by grinding, solubilization using urea, and sterilization. Protein characterization was performed by mass spectrometry. Platelet reactivity was assessed using light transmission aggregometry and flow cytometry. Expression of vascular marker genes was performed in HUVEC and vascular smooth muscle cells (VSMC). The effect of HAM on vascularization was tested in a vascularized micro-organ (VMO) model on a chip.

Results: Native HAM showed a differential protein content compared to Matrigel®, with a significant overrepresentation of proteins involved in cellular motility, adhesion, and endocytosis. Platelet reactivity was limited when compared to Horm® collagen. Cobble-stone and fibroblastic morphology of endothelial cells and VSMC, respectively, cultured on HAM was similar to common culture coatings, however, gene expression analysis resulted in increased expression of VE-Cadherin in endothelial cells as well as increased expression of αSMA and CNN1 and decreased expression of S100A4 in VSMC. In VMO, HAM contributed to an increased vascular diameter compared to fibrin alone.

Conclusion(s): HAM consists of vasculature-specific proteins, which lead to less platelet activity and aggregation. In VSMC, HAM contributes to a more contractile and less synthetic phenotype than control. HUVECs grown on HAM express higher levels of the endothelial marker VE-Cadherin levels, which indicates a tighter endothelial barrier. The results highlight the potential for HAM as an alternative in vitro culture coating which more accurately depicts the in vivo situation.